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Cibotii Rhizoma has a long history of folk usage and clinical applications,which is commonly used to treat symptoms such as waist and knee soreness,numbness of hands and feet.Its chemical components mainly include sugars and glycosides,aromatics,volatile oils,pterosins,flavonoids,steroids,pyranones,amino acids and inorganic elements.Pharmacological studies have shown that Cibotii Rhizoma has antioxidant,anti-inflammatory,anti-osteoporosis,gastrointestinal protection,antibacterial,liver protection,anticancer,analgesic and other effects.This paper systematically reviews the research status of chemical constituents,pharmacological effects and quality control of Cibotii Rhizoma,which provides reference for the development and rational use of Cibotii Rhizoma.
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ObjectiveTo investigate the effect of Fuzheng Huayu prescription on hepatocyte extinction and regeneration in fibrotic liver and its mechanism of action in promoting hepatocyte regeneration. MethodsMice were given intraperitoneal injection of CCl4 for 6 weeks to establish a model of liver cirrhosis， and there were 10 mice in the model group， 10 in the sorafenib group， 10 in the Fuzheng Huayu prescription group， and 9 in the normal control group. Since week 4 of modeling， the mice in the Fuzheng Huayu prescription group and the sorafenib group were given the corresponding drug by gavage at a dose of 4.8 g/kg and 4 mg/kg， respectively， for three consecutive weeks， and those in the normal group and the model group were given an equal volume of sodium carboxymethyl cellulose. Serum liver function parameters were measured； the METAVIR scoring system was used to evaluate liver inflammation and fibrosis stage； Sirius Red staining and hydroxyproline （Hyp） content in liver tissue were used to evaluate collagen deposition； immunohistochemistry was used to measure the protein expression levels of type IV collagen， CD31， CD32b， Ki67， CyclinD1， glutamine synthetase， Wnt2， and HGF， and Western blot was used to measure the expression levels of Wnt2， LRP6， β-catenin， p-β-catenin， and CyclinD1 in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， the Fuzheng Huayu prescription group and the sorafenib group showed the following changes： significant reductions in the serum levels of alanine aminotransferase and aspartate aminotransferase and the content of Hyp in liver tissue （all P<0.01）； a significant reduction in METAVIR score； significant reductions in the expression levels of type Ⅳ collagen and CD31 （all P<0.05） and a significant increase in the expression level of CD32b （P<0.01）； significant reductions in the number of parenchymal extinction lesions and significant increases in the expression levels of Ki67 and CyclinD1 in liver tissue （all P<0.01）； significant increases in the protein expression levels of Wnt2， LRP6， β-catenin， and CyclinD1 and a significant reduction in the protein expression level of p-β-catenin （all P<0.05）； significant increases in the number of cells stained positive for both CD32b and Wnt2. ConclusionFuzheng Huayu prescription can inhibit hepatic sinusoidal capillarization， improve the Wnt2 exocrine function of liver sinusoidal endothelial cells， activate the Wnt/β-catenin signaling pathway associated with hepatocyte regeneration， and finally reverse liver cirrhosis.
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Objective To optimize the extraction process of total phenolic acids from Cibotii rhizoma using Box-Behnken design-response surface methodology and evaluate antioxidant activity of total phenolic acid in vitro.Methods Taking the liquid-solid ratio,ethanol concentration and extraction temperature as influencing factors,and the extraction rate of total phenolic acids as evaluation indicator,the extraction process of total phenolic acids from Cibotii rhizoma was optimized using a three-factor and three-level Box-Behnken design-response surface methodology on the basis of single factor tests.Meanwhile,the scavenging effects of total phenolic acid extract from Cibotii rhizoma on ABTS·+and DPPH· was determined.Results The optimized extraction process for total phenolic acids from Cibotii rhizoma was as follows:the ethanol concentration of 55％,the extraction temperature of 88℃ and the liquid-solid ratio of 60∶1 mL/g.Under these conditions,the extraction rate could reach 8.67％.When the mass concentration of total phenolic acids extract were 2 mg/mL and 1 mg/mL,the clearance rates of ABTS·+and DPPH·were 92.76％and 88.66％,respectively.Conclusion The theoretical values obtained from the response surface optimization method are consistent with the actual measured values,and the extraction process of total phenolic acids from Cibotii rhizoma was simple and feasible.The total phenolic acids extract from Cibotii rhizoma exhibit strong antioxidant activity in vitro.
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Objective To observe the characteristics of C57BL/6N-Tg(1.28HBV)/Vst transgenic hepatitis B virus(HBV-Tg)model mice and analyze their transcriptomic characteristics.Methods Twenty male HBV-Tg mice were divided into an experimental group and a wild-type(control)group(n = 10 mice per group).The virological characteristics of the model mice were evaluated according to serum levels of HBV DNA,HBsAg,and HBeAg,and expression levels of HBsAg and HBcAg in liver tissue.Serum levels of alanine transaminase(ALT)and aspartate transaminase(AST),hematoxylin and eosin(HE)and Sirius red staining,and hydroxyproline(Hyp)in liver tissue were detected to evaluate the degree of liver inflammation and fibrosis.Liver tissue samples were randomly selected from three mice in each group for RNA extraction for high-throughput transcriptome sequencing.Significantly differentially expressed genes were identified using R software.Functional enrichment of differential genes was determined by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses,and genes with significant differences were verified by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results ALT and AST levels were increased in the model group compared with the normal group,with the result for ALT being more significant(P<0.05).HE staining of liver tissue showed enlargement of the liver nucleus and swelling of some hepatocytes in the model group,while Sirius red staining showed a small amount of collagen deposition in the sink area and interlobule in the HBV transgenic group,in the shape of thin lines.A total of 1352 differential genes were obtained by screening(|logFC|>2 and P.adj<0.05),including 703 up-regulated and 649 down-regulated genes.KEGG analysis suggested that differential genes were mainly enriched in the peroxisome proliferator-activated receptor(PPAR)signaling pathway,retinol metabolism,fatty acid degradation,and other pathways(P.adj<0.05).The main significantly up-regulated genes included Cyp4a10,Cyp4a14,Acot1,Acot3,and Ehhadh,and the significantly down-regulated genes included Scn5a、Apol10b、Igddc4、Cxcl1、9530077C05Rik.The trend was consistent after RT-qPCR detection(P<0.05).Conclusions HBV-Tg mice have a tendency to develop spontaneous fibrosis.Transcriptomic analysis showed that chronic hepatitis B mainly involves PPAR signaling,retinol metabolism,fatty acid degradation,drug metabolism,and other pathways.
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Objective:To screen and identify H-2 d-restricted T cell epitopes in fusion (F) and attachment (G) glycoproteins of Nipah virus (NiV) in mice. Methods:The complete peptides (single peptide contains 15 amino acids, and 10 amino acids were repeated in the front and back peptides) derived from F and G antigens were mixed into peptide libraries. BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination. The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design, and spleen cells of immunized mice were collected and analyzed by IFN-γ ELISPOT assay to detect the dominant H-2 d-restricted epitope peptides. Results:Twelve dominant H-2 d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction. Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction. Conclusions:In this study, 12 F antigen-specific and 4 G antigen-specific H-2 d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γ ELISPOT, which could provide reference for immunological analysis of NiV and vaccine research.
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Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.
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Lignin valorization for fuels and value-added products is essential to enhance the profitability and sustainability of biorefineries. Due to the complex and heterogeneous structure of lignin, technical barriers hinder the implementation of economic lignin utilization. Here, we summarize the major challenges facing lignin valorization processes. Different pretreatment methods, especially emerging combinatorial pretreatment approaches for isolating and tailoring lignin are introduced. To overcome the heterogeneity of lignin structure and improve lignin processability, advances in fractionation approaches including organosolv extraction, membrane technology, and gradient precipitation are analyzed and presented. Furthermore, progress in lignin valorization by thermochemical and biological conversion coupling with pretreatment and fractionation are systematically reviewed. Finally, we discuss advanced strategies and perspectives for future research involving biomass pretreatment, lignin fractionation and conversion processes.
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Biomasa , LigninaRESUMEN
Chikungunya virus(CHIKV)is a member of the genus Alphavirus, family Togaviridae and transmitted to humans by Aedes aegypti and Aedes albopictus. It mainly causes Chikungunya fever (CHIKF) with high fever, rash and multiple arthritis/joint pain as the main clinical symptoms, and is a major public health threat of global concern. There is still no vaccine or effective antiviral drug against CHIKV. This review briefly summarized the genomic and epidemiological characteristics of CHIKV, features of immune response to major CHIKV antigens and latest progress in vaccine development.
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Liver fibrosis is commonly seen in various chronic liver diseases. In history, traditional Chinese medicine (TCM) clinical practice based on syndrome differentiation and treatment formed unique experience in the diagnosis and treatment of liver fibrosis; in recent years, studies based on experimental exploration and evidence-based medicine have formed effective Chinese patent drugs and experienced prescriptions for the treatment of liver fibrosis, as well as integrated traditional Chinese and Western medicine regimens. With the improvement in the quality of TCM drugs and the development of related clinical studies, new TCM drugs with stable quality and marked clinical effect will be launched for the treatment of liver fibrosis, so as to meet the clinical needs of patients with chronic liver diseases and promote the modern and international development of TCM.
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Hepatitis B virus (HBV)/hepatitis D virus (HDV) co-infection can accelerate the progression of liver diseases, but due to the limitations of the detection methods for HDV RNA, the epidemiology of HDV remains unclear. Further studies are needed to investigate the pathogenesis of HBV/HDV co-infection caused by HDV and related treatment methods. By reviewing the epidemiological features of HDV, HDV-specific immunity, genotype, and pathogenesis in China and other countries and summarizing HDV detection methods and treatment methods, this article shows that current detection methods for HDV infection have low sensitivity and specificity, with a lack of standardized detection methods. Exploration of new HDV detection methods will help to develop new and effective anti-HDV drugs, which has great significance in improving the outcome of HDV infection.
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Hepatitis B virus (HBV)/hepatitis D virus (HDV) co-infection can accelerate the progression of liver diseases, but due to the limitations of the detection methods for HDV RNA, the epidemiology of HDV remains unclear. Further studies are needed to investigate the pathogenesis of HBV/HDV co-infection caused by HDV and related treatment methods. By reviewing the epidemiological features of HDV, HDV-specific immunity, genotype, and pathogenesis in China and other countries and summarizing HDV detection methods and treatment methods, this article shows that current detection methods for HDV infection have low sensitivity and specificity, with a lack of standardized detection methods. Exploration of new HDV detection methods will help to develop new and effective anti-HDV drugs, which has great significance in improving the outcome of HDV infection.
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ObjectiveTo investigate the changes in positive staining of CD34, CK7, and CK19 and amount of fibrous collagen deposition in patients with chronic hepatitis B (CHB) and the pathological basis affecting FibroTouch measurements. MethodsA retrospective analysis was performed for the clinical data of 72 CHB patients who visited Department of Liver Cirrhosis in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2015 to December 2017. The amount of positive immunohistochemical staining of CD34, CK7, and CK19 was calculated, as well as the amount of fibrous collagen deposition in Masson trichrome staining and liver stiffness measurement (LSM) by FibroTouch. The t-test was used for comparison of normally distributed continuous data between two groups. The Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data or continuous data with heterogeneity of variance between two groups. The chi-square test was used for comparison of categorical data between groups, and the Kruskal-Wallis H test was used for comparison of ranked data between multiple groups. The receiver operating characteristic (ROC) curve was used to analyze the value of LSM in the diagnosis of hepatitis B cirrhosis, and the logistic regression model was used for multivariate analysis. ResultsWith the increase in inflammation degree, there was no significant change in the amount of positive staining of CD34 (P＞0.05), while there were significant increases in the amount of positive staining of CK19 and the amount of fibrous collagen deposition (H=9.02 and 14.12, P=0011 and 0.001). With the progression of liver fibrosis, there were significant increases in the amount of positive staining of CD34 and CK7 and the amount of fibrous collagen deposition (H=10.26, 16.29, and 22.97, P=0.016, 0.001, and ＜0.001). The logistic regression analysis showed that the amount of positive staining of CK7 (Wald=4.756, P=0.029) and the amount of fibrous collagen deposition (Wald=4.757, P=0.029) were independent influencing factors for FibroTouch measurements. ConclusionIncreases in the amount of fibrous collagen deposition and the amount of positive staining of CK7 may lead to increased FibroTouch measurements.
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Objective@#To investigate the correlation of liver stiffness measured by FibroTouch (FT) and FibroScan (FS) with Ishak fibrosis score in patients with chronic hepatitis B.@*Methods@#A total of 313 patients with chronic hepatitis B who visited Department of Liver Cirrhosis in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2014 to May 2016 were enrolled. All the patients underwent liver biopsy, and FT and FS were used to determine liver stiffness measurement (LSM). Serum biochemical parameters were measured, and the aspartate aminotransferase-to-platelet ratio index (APRI) in a multi-parameter model of liver fibrosis and fibrosis-4 (FIB-4) index were calculated. The consistency between the results of four noninvasive examinations and Ishak fibrosis score was compared. The t-test was used for comparison of LSM determined by FT and FS. Pearson correlation analysis was used investigate the correlation between LSM determined by FT and FS; Spearman correlation analysis was used to investigate the correlation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and Knodell score with LSM determined by FT and FS; the correlation between LSM determined by FT and FS and fibrosis stage was analyzed by partial correlation analysis adjusted by Knodell score for liver inflammatory activity; Spearman correlation analysis was used for APRI, FIB-4, and fibrosis stage. Based on the Ishak fibrosis score, the receiver operating characteristic (ROC) curve was used to analyze the values of four noninvasive methods in the diagnosis of liver fibrosis.@*Results@#There was no significant difference in LSM measured by FT and FS in all patients (15.75±9.42 kPa vs 15.42±10.52 kPa, P > 0.05) and Pearson correlation analysis indicated a significant positive correlation between them (r = 0.858, P < 0.01); serum ALT and AST levels and liver inflammatory activity were correlated with LSM determined by FT and FS. There was a significant positive correlation between LSM determined by FT and FS and fibrosis stage (r = 0.501 and 0.526, both P < 0.001), and APRI and FIB-4 were also positively correlated with fibrosis stage (r = 0.236 and 0.218, both P < 0.001). Based on the Ishak fibrosis score, in the diagnosis of fibrosis stages F3, F4, F5, and F6, the areas under the ROC curve were 0.915/0.856/0.839/0.816 for FT, 0.933/0.883/0.849/0.856 for FS, 0.618/0.630/0.608/0.638 for APRI, and 0.614/0.624/0.595/0.649 for FIB-4, and FT and FS had a significantly larger areas under the ROC curve than APRI and FIB-4.@*Conclusion@#LSM determined by FT or FS has a good correlation with the Ishak fibrosis score, so FT and FS have a significantly better diagnostic performance for liver fibrosis than APRI and FIB-4.
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Objective The number of children diagnosed with autism spectrum disorder (ASD)has increased dramatically in the past years.Parents of children with ASD usually experience a variety of chronic and acute stressors that may decrease marital satisfaction and family functioning.The aim was to explore the characteristics of parental per-sonality,mood disorders and family functions of the Children with ASD and to explore the relevance factors in order to draw out methods to improve parental family functions.Methods A cross -sectional survey was conducted and 593 outpatients with ASD diagnosed in Shanghai Mental Health Center,Shanghai Jiao Tong University School of Medicine form January 201 0 to January 201 4 were straight into the group,their parents were tested by Family Assessment Device (FAD),Zung Self -Rating Depression Scale(SDS),Zung Self -Rating Anxiety Scale(SAS)and Eysenck Personality Questionnaire(EPQ).SPSS 1 7.0 software was adopted to establish the database and statistical analysis.Results Ac-cording to FAD,parents of children with ASD had lower scores in problem solving[(2.1 0 ±0.37)scores]and roles [(2.09 ±0.27)scores](t =-6.669,P =0.000;t =-1 8.707,P =0.000)than the standard scores,while other di-mensions score were significantly higher than the standard scores in communicdtion[(2.25 ±0.35)scores],emotional reaction[(2.38 ±0.40)scores],emotional involvement[(2.36 ±0.37)scores],behavioral control[(2.31 ±0.27) scores]and general function[(2.1 2 ±0.38)scores],there were significant differences(t =3.1 35,1 1 .1 1 5,1 6.997, 37.1 1 6,8.064;P =0.002,0.000,0.000,0.000,0.000).For EPQ of parents,mother had obviously higher scores than father in psychoticism scale[(53.68 ±1 .36)scores vs (52.65 ±1 .1 7)scores,t =-1 3.841 ,P =0.000],and signifi-cantly lower scores in Lie scale[(52.27 ±0.79)scores vs (52.1 7 ±0.75)scores,t =2.449,P =0.01 4].Compared to fathers,mothers were coded as higher depression and anxiety scores(t =-6.276,-6.440,all P =0.000)and higher prevalence rate(χ2 =1 5.893,27.592,all P =0.000),which were statistically significant.According to the asso-ciation study of FAD and the others(especially in mood of parents),it was found that several dimensions showed obvious correlation.Conclusions Parents of children with ASD have family function damaged badly,except for prob-lem solving and roles.The parents showed different personality traits and mood disorders,particularly mothers′mood disorder problem was prominent.Parental family functioning were closely related to their personality traits and emotional disorders in various dimensionalities,which was worthy of attention.
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Objective To establish the reference value ranges of specific thyroid function indexes among pregnant women in the Longgang District Maternity and Child Healthcare Hospital for avoiding the misdiagnosis and missed diagnosis of pregnant thyroid diseases .Methods A total of 637 normal pregnant women with establishing the card in the outpatient department of this hospital and 201 non‐pregnant women undergoing the normal physical .The fasting venous blood was collected .The values of FT3 ,FT4 ,T3 , T4 and TSH were detected by adopting the chemiluminescence immunoassay .The median and 95% credibility interval statistical method was used to establish the normal reference value ranges of specific serum free triiodothyronine(FT3) ,free thyroxine(FT4) , three icdine thyroid gland(T3) ,total thyroid hormone(T4) and thyroid stimulating hormone(TSH) in pregnancy .Results The me‐dian(M value) and double‐side limiting value(P2 .5 and P97 .5 ) were used to express the reference value ranges of thyroid hormones in early ,middle and late pregnant women and non‐pregnant women .FT3 and FT4 were significantly increased at early pregnancy ,then gradually increased with the pregnant period progression ,began to decrease from middle pregnancy and were higher than the levels before pregnancy .FT3 and FT4 were slightly increased ,then progressively decreased .TSH was significantly decreased in early pregnancy ,began to increase at middle pregnancy and was higher than the level in non‐pregnant women until late pregnancy .Conclu‐sion The establishment of reference value ranges of specific thyroid hormones during various stages of pregnancy ,which provides the clinical data for the diagnosis and treatment of thyroid disease during pregnant period;the change rule of thyroid hormones in pregnant women provides a theoretical foundation for the prevention of thyroid disease during pregnant period .
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The advances of industrial biotechnology highly depend on the development of industrial bioprocess researches. In China, we are facing several challenges because of a huge national industrial fermentation capacity. The industrial bioprocess development experienced several main stages. This work mainly reviews the development of the industrial bioprocess in China during the past 30 or 40 years: including the early stage kinetics model study derived from classical chemical engineering, researching method based on control theory, multiple-parameter analysis techniques of on-line measuring instruments and techniques, and multi-scale analysis theory, and also solid state fermentation techniques and fermenters. In addition, the cutting edge of bioprocess engineering was also addressed.
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Bioingeniería , Historia , Reactores Biológicos , Biotecnología , Ingeniería Química , China , Fermentación , Historia del Siglo XX , Historia del Siglo XXIRESUMEN
<p><b>OBJECTIVE</b>To investigate the methods for assessing angiogenesis in experimental liver fibrosis.</p><p><b>METHODS</b>Male C57BL/6 mice were randomly divided into a normal (unmodeled) group and a liver fibrosis model group.The model was established by a 4-week course of 10% CCl4 solution (in olive oil) intraperitoneal injection. Liver vasculature was observed by magnetic resonance (MR),computed tomography (CT),synchrotron radiation X-ray,and von Willibrand factor(vWF) immunofluorescence staining. Liver inflammation and fibrosis were observed by staining with hematoxylin-eosin and Sirius red.The t-test and Pearson's correlation analysis were used for statistical analyses.</p><p><b>RESULTS</b>Compared with the normal group,the model group had more robust inflammation and fibrosis in liver tissue.The liver tissue in the modeled mice showed significant deceases in MR signal intensity on T2WI before and after enhancement (386.67+/-69.04 vs.492.67+/-112.50,t =-2.456, P=0.026).The liver tissue in the modeled mice also showed significantly decreased enhancement CT values (P < 0.01).Synchrotron radiation X-ray imaging showed that the small vessels in the liver tissues of the modeled mice were significantly increased compared to that in the normal mice (P < 0.01).The MR T2W enhanced signal value (439.67+/-104.80, Pearson's r=0.714, P=0.009) and microvascular relative length (676.53+/-122.75, Pearson's r=0.791, P=0.002) were positive correlated with microvessel density (MVD) (14.50+/-5.95),as shown by detection of labeled vWF.The before and after CT enhancement difference (132.60+/-57.02, Pearson's r=-0.612, P=0.034) was negatively correlated with MVD.</p><p><b>CONCLUSION</b>s MR,CT and synchrotron radiation X-ray imaging can be used for assessing angiogenesis in liver fibrosis,and the findings from each are correlated with the traditional MVD detection method. The two-dimensional imaging of synchrotron radiation X-ray is more intuitive,and has been confirmed as an effective evaluation method for liver angiogenesis.</p>
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Animales , Masculino , Ratones , Modelos Animales de Enfermedad , Inflamación , Patología , Cirrosis Hepática , Patología , Pruebas de Función Hepática , Espectroscopía de Resonancia Magnética , Ratones Endogámicos C57BL , Neovascularización Patológica , Sincrotrones , Tomografía Computarizada por Rayos XRESUMEN
Objective To observe the therapeutically effect of kangnao liquid on Pi3k mRNA and Aktm RNA ex-pressions in rats with focal cerebral ischemia-reperfusion (I/R) injury. Methods 180 male SD rats were randomly divided into 6 groups:sham operated group, model group, three kangnao liquid groups (high-dose, medium-dose and low-dose) and nimodipine group. Rats in kangnao liquid groups were administrated with kangnao liquid of 24 g/(kg · d), 12 g/(kg · d) and 6 g/(kg · d), orally once a day. Rats in nimodipine group were given nimodipine 1 mg/(kg · d). Rats in model group and sham group were treated with the same volume of distilled water for 7 days. The animal model of middle cerebral artery occlusion (MCAO) was established by a monofilament method from right internal carotid artery. The neurological evaluation was per-formed 24 h after reperfusion. The in situ hybridization was used to investigate the expression levels of Pi3k mRNA and Akt mRNA in rats on 12 h, 24 h, 48 h, 72 h and 168 h after ischemia for 2 h. Results Compared with model group, neurological functions were improved significantly in kangnao liquid groups. The expression levels of Pi3k mRNA and Akt mRNA were al-so significantly higher in kangnao liquid groups than those of model group. The expression levels of Pi3k mRNA and Akt mRNA were significantly higher in nimodipine group than those of model group, but which were lower compared with those of high-dose and medium-dose kangnao liquid groups. Conclusion Kangnao liquid can protect nerve cells by enhancing the expressions of Pi3k mRNA and Akt mRNA in rats with cerebral ischemia-reprefusion injury.
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<p><b>OBJECTIVE</b>To imitate tumor microenvironment in vivo through construction of two-layer cell spheroid as a three-dimensional tumor model, and to validate its application in the study of Cx43 expression in colorectal cancer cell-fibroblast interactions and colorectal cancer progression.</p><p><b>METHODS</b>The two-layer cell spheroid was constructed from SW620 colorectal cancer cells and HELF fibroblasts. The expression of Cx43 in the spheroid was detected by immunocytochemistry. The expression of Cx43 in cultured SW620 cells with or without co-cultured fibroblasts was detected by immunocytochemistry and immunofluorescence. The expression of Cx43 in colorectal cancer tissue was detected by immunohistochemistry.</p><p><b>RESULTS</b>The spheroid showed well-defined cellular morphology and clear boundary between two cell lines.Significant expression of Cx43 was found along the boundary.SW620 cells had no expression of Cx43 when cultured alone, while the expression of Cx43 was induced upon co-culturing with fibroblasts.In the colorectal cancer tissue, expression of Cx43 was minimal in the centre of tumor in contrast to an upregulated expression at invasive front.</p><p><b>CONCLUSIONS</b>The two-layer cell spheroid is an observable and sensitive model for cell-cell interaction for studies of tumor microenvironment.It can simulate colorectal cancer cell-fibroblast interactions through up-regulation of Cx43 expression.</p>
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Humanos , Comunicación Celular , Línea Celular Tumoral , Técnicas de Cocultivo , Neoplasias Colorrectales , Metabolismo , Patología , Conexina 43 , Metabolismo , Fibroblastos , Biología Celular , Esferoides Celulares , Biología Celular , Metabolismo , Microambiente Tumoral , Regulación hacia ArribaRESUMEN
<p><b>OBJECTIVE</b>To observe the effect of traditional Chinese medicine formulas clearing away heat and promoting blood circulation-Biejiayinzi (BJYZ), Gexiazhuyu Tang (GXZYT) and Fugan Wan (FGW) on liver fibrosis in CCl4 mice by screening and analyzing formula-syndrome database of kidney and liver fibrosis based on the principle of formula-syndrome, compared with pivot-harmonizing decoction.</p><p><b>METHOD</b>Ten-week-old male C57BL/6 mice, with the weight of (20 +/- 3) g, were randomly divided into 6 groups: the normal group, the model group, the BJYZ group, the GXZY group, the FGW group and the XST group. Except the normal group, other groups were abdominally injected with 10% CCl4 olive oil solution a dose of 2 mL x kg(-1) body weight for four weeks, three times each week. Meanwhile, the latter four groups were administered with extracts of BJYZ, GXZYT, FGW and XST, respectively, once every day, concomitantly continued CCl4 administration. The normal and the model groups were given the same volume of deionized water. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (gamma-GT), Alb and TBil were detected by chemiluminescence. The hydroxyproline (HYP) content was detected by acid hydrolysis method. The hepatic collagen deposition was evaluated with Sirius red staining.</p><p><b>RESULT</b>Compared with the normal group, the model group recorded notable decrease in weight and increase in the ratio of liver weight and body weight and the ratio of spleen weight and body weight, with obvious fatty degeneration and inflammatory necrosis in liver cells. Collagen fiber deposition was so notable to form fibrous septums and pseudolobules. The levels of serum ALT, AST, TBil, gamma-GT, the HYP content in liver tissue and the deposition of hepatic collagen were significantly increased in the model group. Compared with model group, Serum AST were significantly decreased in BJYZ group as gamma-GT decreased in the GXZYT group, without notable decrease in degeneration and inflammatory necrosis in liver cells and collagen deposition. The GXZYT group showed significant decrease in gamma-GT, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The ratio of liver weight and body weight, AST, gamma-GT and HYP content were significantly decreased in the FGW group, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The XST group showed decrease in the ratio of liver weight and body weight, with no obvious change in inflammation and fibrosis of hepatic tissue.</p><p><b>CONCLUSION</b>FGW shows the best effect of prevention and treatment of liver fibrosis in CCl4 mice.</p>